Transplantation of stromal cells transduced with the human IL3 gene to stimulate hematopoiesis in human fetal bone grafts in non-obese, diabetic-severe combined immunodeficiency mice

Abstract

The non-obese diabetic-severe combined immunodeficiency (NOD-SCID) mouse is a convenient host for human hematopoietic tissues and cells. Human fetal bone fragments engrafted subcutaneously in NOD-SCID mice sustain human hematopoiesis for several months. MS5 murine bone marrow stromal cells were transfected by electroporation with a plasmid containing the human interleukin-3 gene. As expected, stably transfected hu-IL3-MS5 cells supported human hematopoiesis in vitro more efficiently than MS5 cells. hu-IL3-MS5 cells were then injected intravenously into hu-NOD-SCID mice to test their ability to home to the mouse and/or human bone marrow, and to evaluate the role of hu-IL3 secretion on human hematopoiesis in vivo. hu-IL3 was detected in the mouse serum for up to an observation time of 8 weeks. hu-IL3-MS5 cells engrafted the bone marrow, spleen, liver and lungs of the mice but also the human bone graft. The presence of hu-IL3-MS5 cells in the human bone significantly stimulated local human hematopoiesis. This setting could be used to model the bone marrow homing of intravenously injected stromal cells or stromal cell precursors. The same experimental principle could also be applied in a therapeutic perspective to malignant human bone marrow hematopoiesis.

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

“…Engraftment and persistence of human MSCs expressing human factor VIII, human factor IX, or human IL-3 was demonstrated in unconditioned immunodeficient mice following transplantation. 40,42,43,70,71 Similar experiments with human transgene products have been carried out in immunocompetent transplant models. For example, gene-modified canine MSCs expressing human growth hormone or human factor IX were infused in unconditioned normal dogs.…”

“…29,[37][38][39][40][41] Various studies have shown the delivery of a secretable human transgene product by MSCs administered via the intravenous route in immunodeficient rodents. 40,42,43 When infused intravenously in immunocompetent recipients, genetically engineered MSCs engendered transiently detectable plasma levels of a species mismatched secreted protein. 44,45 Therefore, the successful utilization of MSCs in transgenic adoptive cell therapy will depend upon their demonstrated ability to engraft in nonmyeloablated, immunocompetent recipients and lead to a sustained pharmacological effect.…”

Regulation of hematopoietic stem cell fate

“…Therefore, there is plenty of opportunity for cell-cell signaling. Studies in which HSC were expanded in vitro have shown that contact with stromal cells enhances HSC proliferation (Brouard et al, 1998;Dorshkind, 1990). HSCs receive messages from adjacent cells via gap junctions (connexins are expressed by developing hematopoietic cells (for recent reviews see MontecinoRodriguez and Dorshkind, 2001;Rosendaal and Krenacs, 2000) and receptor/ligand interactions between adjacent cells.…”

Homing of in vitro expanded Stro-1- or Stro-1+ human mesenchymal stem cells into the NOD/SCID mouse and their role in supporting human CD34 cell engraftment

“…Recent results have shown that cotransplantation of human ex vivo-expanded mesenchymal stem cells (MSCs) together with hematopoietic stem cells hastens hematopoietic recovery following a bone marrow (BM) transplantation in animal models [1][2][3][4] and in humans. [5][6][7] Human BM contains 2 cell compartments, the hematopoietic cell compartment and the stromal cell compartment, which constitute MSCs.…”